AWWA QTC98264

AWWA QTC98264 "Grab Sample PCR" for the Detection of Viruses in Ground Water Samples

Conference Proceeding by American Water Works Association, 01/01/1998

Abbaszadegan, Morteza; Denhart, Mark

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The objective of this work was to develop a RT-PCR based low cost, highly sensitive system to detect viruses in a one liter sample volume, to be used in the routine monitoring of viral contamination of water sources intended for drinking. A rapid method for the detection of Poliovirus type 1, MS2, and PRD-1 has been developed using a small, low cost 1MDS capsule filter. One liter of water was passed through the filter at a rate of 100 mL per minute. Virus elution from the filter was accomplished using 10 to 15 mL of 1.5% beef extract solution. To concentrate recovered viruses, the filter eluent was reduced in volume to less than 150 uL using a high molecular weight cutoff spin column. Poliovirus RNA was extracted from the concentrates and reduced in volume to 20 uL and a portion used for RT-PCR assay. The sample collection, concentration, and extraction method can detect as little as 1 PFU of virus in one liter of water. The sample collection and elution is an improvement over the currently used large volume method in terms of cost, time, labor and sensitivity per liter.

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